FIGURE 6.

Transgenic NA protein produced during development persists into adulthood. Western blot analyses of NA and UNC79 expression from head extracts of Drosophila males raised at 19°C or 29°C and maintained at 19°C or 29°C as adults, as indicated. (A) Representative Western blot data. Lane 1 (na+): elavGAL4 na⁺; tubGAL80^ts/ + males, raised and maintained at 19°C. Lanes 2–6: elavGAL4 na^har; tubGAL80^ts/ +; UAS-na/+ males, with developmental (dev) and adult temperature conditions indicated below each lane (dev temperature – adult temperature). For NA (top blot), two bands are typically observed for the endogenous protein (lane 1) while transgenic UAS-na produces a single band of lower molecular weight (lanes 2–6) (Lear et al., 2005). (B) Quantitation of NA and UNC79 levels for genotypes and conditions in panel (A) (see key). Bars indicate normalized protein levels, as a percentage of levels observed in elavGAL4 na^har; tubGAL80^ts/ +; UAS-na/+ flies in the 29°C development- 29°C adult condition (red bars; part A, lane 3). Error bars indicate standard error of the mean, as determined from two independent experiments. For elavGAL4 na^har; tubGAL80^ts/ +; UAS-na/+ flies raised and maintained at 19°C (light blue bars; panel A, lane 2), asterisks denote significant differences from elavGAL4; tubGAL80^ts/ + flies maintained in the same conditions (white bars, ^∗p < 0.05, ^∗∗p < 0.01; panel A, lane 1). For elavGAL4 na^har; tubGAL80^ts/ +; UAS-na/+ flies raised at 29°C, adults were either maintained at 29°C (red bars; panel A, lane 3) or were shifted to 19°C for 1–2 days (pink bars; panel A, lane 4), 3–4 days (purple bars; panel A, lane 5), or 5–6 days (dark blue bars; panel A, lane 6). Protein levels in the 29°C to 19°C shifted conditions were not significantly different from the 29°C to 29°C control (n.s.; p > 0.08). Statistical comparisons were made using Student’s t-test.